Lipoamide Dehydrogenase/DLD Rabbit Polyclonal Antibody, Unconjugated

Artikelnummer: BYT-ORB312105
Artikelname: Lipoamide Dehydrogenase/DLD Rabbit Polyclonal Antibody, Unconjugated
Artikelnummer: BYT-ORB312105
Hersteller Artikelnummer: orb312105
Alternativnummer: BYT-ORB312105-100
Hersteller: Biorbyt
Wirt: Rabbit
Kategorie: Antikörper
Applikation: FC, ICC, IF, IHC, WB
Spezies Reaktivität: Human, Mouse, Rat
Immunogen: E.coli-derived human DLD recombinant protein (Position: K300-F509). Human DLD shares 96.2% and 95.7% amino acid (aa) sequence identity with mouse and rat DLD, respectively.
Konjugation: Unconjugated
Alternative Synonym: Dihydrolipoyl dehydrogenase, mitochondrial, 1.8.1.4, Dihydrolipoamide dehydrogenase, Glycine cleavage system L protein, DLD, GCSL, LAD, PHE3
Lipoamide Dehydrogenase/DLD Rabbit Polyclonal Antibody
Klonalität: Polyclonal
Konzentration: Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.
Molekulargewicht: 54 kDa
UniProt: P09622
Puffer: Each vial contains antibody formulated with stabilizing components, 0.9 mg NaCl, 0.2 mg Na2HPO4, and 0.05 mg NaN3. *This antibody is supplied in a stabilized formulation. Compatibility with conjugation reactions depends on the chemistry of the conjugation
Formulierung: Lyophilized
Target-Kategorie: Dihydrolipoyl dehydrogenase, mitochondrial
Application Verdünnung: Immunohistochemistry (Paraffin-embedded Section), 0.5-1µg/ml, Human Western blot, 0.1-0.5µg/ml, Human, Mouse, Rat Immunocytochemistry/Immunofluorescence, 2µg/ml, Human Flow Cytometry (Fixed), 1-3µg/1x10 6 cells, Human
Flow Cytometry analysis of A549 cells using anti-DLD antibody. Overlay histogram showing A549 cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-DLD Antibody (1 µg/1x10 6 cells) for 30 min at 20C. DyLight488 conjugated goat anti-rabbit IgG (5-10 µg/1x10 6 cells) was used as secondary antibody for 30 minutes at 20C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10 6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
WB analysis of DLD using anti-DLD antibody.Lane 1:Rat Brain Tissue,2:
IF analysis of DLD using anti-DLD antibody. DLD was detected in immunocytochemical section of MCF-7 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2 µg/mL rabbit anti-DLD Antibody overnight at 4C. DyLight488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
IHC analysis of DLD using anti-DLD antibody. DLD was detected in a paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-DLD Antibody overnight at 4C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
Western blot analysis of DLD using anti-DLD antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human K562 whole cell lysates, Lane 2: human A431 whole cell lysates, Lane 3: human SiHa whole cell lysates, Lane 4: human U251 whole cell lysates, Lane 5: rat liver tissue lysates, Lane 6: rat heart tissue lysates, Lane 7: mouse liver tissue lysates, Lane 8: mouse heart tissue lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-DLD antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-DyLight 647 Conjugated secondary antibody at a dilution of 1:2000 for 1.5 hour at RT. A specific band was detected for DLD at approximately 54 kDa. The expected band size for DLD is at 54 kDa.
Western blot analysis of DLD using anti-DLD antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human K562 whole cell lysates, Lane 2: human A431 whole cell lysates, Lane 3: human SiHa whole cell lysates, Lane 4: human U251 whole cell lysates, Lane 5: rat liver tissue lysates, Lane 6: rat heart tissue lysates, Lane 7: mouse liver tissue lysates, Lane 8: mouse heart tissue lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-DLD antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a